[Histonet] Tissue marking dyes
I am wanting to 'cut' the dense tissue dye(s) by TBS (probably same as Cancer Diagnostics or any other). Orange is very thick, so is green. I'm being a bit lazy here, but I was wondering if anyone else is going this? Do you use water, alcohol or H2O2? Any direction would be helpful. Thanks - Bill William (Bill) O'Donnell, HT (ASCP) QIHC Senior Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 SERENITY is not freedom from the storm, but peace amid the storm. Cultivate it in PRAYER! This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Histology Assistant Job Description
Happy Wednesday Everyone, Does anyone have a histology assistant job description that they could share with me? Thanks in advance for your help, Amy Self Histology Lab Senior Tech Lab Georgetown Memorial Hospital 606 Black River Road Georgetown, SC 29440 843-520-8711 as...@georgetownhospitalsystem.org NOTE: The information contained in this message may be privileged, confidential and protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to this message and deleting it from your computer. Thank you. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] manual IHC staining
Hi all- Does anyone out there still do manual, offline IHC staining? We currently do about 20,000 IHC slides per year and my boss would like to consider dropping the automatic stainers and doing them by hand. If you do manual staining, how long does it take, how much tech time is needed, and what about CAP inspections? Thanks in advance, Mehndi Helgren Dominion Pathology Laboratories 733 Boush St. Suite 200 Norfolk, VA 23510 757-664-7901 hi...@pathlab.us ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Old equipment
Would anyone be interested in purchasing some old equipment? We have 2 Ventana Benchmarks that will be available in late November/ early December and a Leica RM 2155 microtome. The Benchmarks work, my company just upgraded to the Benchmark XT. The Leica microtome was having some issues but we couldn't get service on it so we bought a new one. Thank you Fawn Bomar - This electronic message may contain information that is confidential or legally privileged. It is intended only for the use of the individual(s) and entity named as recipients in the message. If you are not an intended recipient of this message, please notify the sender immediately and delete the material from any computer. Do not deliver, distribute, or copy this message, and do not disclose its contents or take any action in reliance on the information it contains. Thank you ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
Re: [Histonet] manual IHC staining
There is no problem with CAP inspections if you do them manually. The tech time is the same for HIER, but it increases for the actual procedure. Please go to http://www.histosearch.com/rene.html and you will find times for manual IHC Generally speaking it will be around 10 mins. per slide per procedure. René J. From: Histology hi...@pathlab.us To: histonet@lists.utsouthwestern.edu Sent: Wednesday, October 16, 2013 12:46 PM Subject: [Histonet] manual IHC staining Hi all- Does anyone out there still do manual, offline IHC staining? We currently do about 20,000 IHC slides per year and my boss would like to consider dropping the automatic stainers and doing them by hand. If you do manual staining, how long does it take, how much tech time is needed, and what about CAP inspections? Thanks in advance, Mehndi Helgren Dominion Pathology Laboratories 733 Boush St. Suite 200 Norfolk, VA 23510 757-664-7901 hi...@pathlab.us ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Re: Histonet Digest, Vol 119, Issue 20
Hello Allyse, Some questions to your question: Date: Tue, 15 Oct 2013 14:43:32 -0400 From: Allyse Mazzarelli allyse...@gmail.com Subject: [Histonet] Autofluorescence on Retina Tissue I've consistently run immunofluorescence on pig retina and I seem to have a severe case of autofluorescence/background. How are you fixing the tissue? Are you using NH4Cl, 0.1 M glycine etc... to quench auto-fluorescence? Where do you see the auto-florescence? Which layers of the retina? Do you have a negative control slides (no primaries or secondaries as well as no primaries with secondaries)? What do they look like? What type of microscope are you using? Confocal? Are you using a nuclear stain such as hoechst? In my experience there is normally a lot of auto-fluorescence in the retina to start with. Fixation tends to make this worse... Using 0.1 M glycine helped me a lot in reducing this problem but the retinal pigment epithelium layer and the chorion behind it would still auto-fluoresce terribly. The former is due to lipofuscin so maybe a H2O2 blocking step may take care of it... Anyone tried? But now I digress... Unless you have negative controls you don't know what the source of the auto-fluorescence is. Do you have controls, what do they look like? I was lucky, as I was working with a confocal and could tune out a lot of the background fluorescence. If you work with a normal fluorescence microscope you may have issues, especially if you have tick sections. IF you are using hoechst as a counter stain for nuclei you may be using too much of the dye. I had to use as little as 0.5 ug/mL for fixed section before I stopped getting bleed-through into other channels from the Hoechst. It showed up in the green and red channels. Hope this helps... -- Tyrone Genade http://tgenade.freeshell.org email: tgen...@gmail.com tel: +27-84-632-1925 (c) Romans 6:23: The gift of God is eternal life through Christ Jesus our Lord. To find out how to receive this FREE gift visit http://www.alpha.org. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: manual IHC staining
Wow - I can't imagine wanting to go to manual staining for 20,000 slides per year. That's about 80-100 per day. We used to do manual staining but only about 20-40 slides per day. Unless you are running a very limited panel of antibodies, you will be hopping to keep up with all the steps. I wouldn't think you would be batching all slides in one run, so you will have timers going off all the time. You or your techs will not have time to do anything else but stain, wash, apply reagents, wash, etc. You might want to do some trial runs for timing, work flows etc before you make the decision to ditch the autostainer. I would also consider consistency of staining , correct antibodies being added to the correct slides, and amount of reagents (no matter how careful you are, you will put more on than the stainer). Just my thoughts. We do 30-35,000 slides per year, we have three Ventana Ultras and they are busy all the time. and so are my 2 techs. Sue Sue Hunter, Supervisor Advanced Diagnostics Beaumont Health System Royal Oak MI 248-898-5146 shun...@beaumont.edu -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Histology Sent: Wednesday, October 16, 2013 12:47 PM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] manual IHC staining Hi all- Does anyone out there still do manual, offline IHC staining? We currently do about 20,000 IHC slides per year and my boss would like to consider dropping the automatic stainers and doing them by hand. If you do manual staining, how long does it take, how much tech time is needed, and what about CAP inspections? Thanks in advance, Mehndi Helgren Dominion Pathology Laboratories 733 Boush St. Suite 200 Norfolk, VA 23510 757-664-7901 hi...@pathlab.us ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: manual IHC staining
Mehndi, Why? Is your automation costing a lot? Your average is 76 slides/day. That is a substantial number. If your automation is costing a lot, then look for a less costly alternative, like the BioCare or Thermo Fisher (Lab Vision) instruments. Those are completely open systems that can have very low costs if you are careful choosing reagents. We do maybe 50 IF slides per day in our kidney lab and would not get rid of automation. In fact, I would only get rid of automation if we were batching slides only a couple days per week. It saves so much hands on time that only if you have extra staff and very cheap labor costs is it worth dropping. My cost calculations show that even at only 50 slides per day we spend only 10% of our total cost of the test on the Dako (Autostainer Plus, same as Lab Vision 480) automation portion (including service contract costs). The major factor is the cost of reagents. And reagent cost won't change much with manual unless you change all your reagents. But your labor costs will skyrocket. I recommend keeping automation, but evaluate your platform for excess costs. Tim Morken UCSF Pathology -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of Histology Sent: Wednesday, October 16, 2013 9:47 AM To: histonet@lists.utsouthwestern.edu Subject: [Histonet] manual IHC staining Hi all- Does anyone out there still do manual, offline IHC staining? We currently do about 20,000 IHC slides per year and my boss would like to consider dropping the automatic stainers and doing them by hand. If you do manual staining, how long does it take, how much tech time is needed, and what about CAP inspections? Thanks in advance, Mehndi Helgren Dominion Pathology Laboratories 733 Boush St. Suite 200 Norfolk, VA 23510 757-664-7901 hi...@pathlab.us ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
RE: [Histonet] Autofluorescence on Retina Tissue
Not sure if this will help with your retinas but we use the copper sulfate treatment on all our fluorescent stains and it eliminates autofluorescence in the RBC completely and helps with other types of autofluorescence. The treatment to be use depends on the cause of the autofluorescence. We counter stain our slides with DAPI. Here are all of our present autofluorescent treatments. Autofluorescence Quenching 1) The best ways to address the issue of autofluorescence in order of preference: a) Avoid it i) Not always possible. b) Try to filter it out during image acquisition i) Difficult due to the broad emission spectrum. c) Chemically remove it i) Can also reduce real signal. 2) 10 mM Copper Sulfate a) This treatment is primarily for inhibition of autofluorescence in Red Blood cells, but helps decrease autofluorescence in connective tissue. b) 10 mM Copper Sulfate i) Cupric Sulfate...1.25 gm. ii) 50 mM Ammonium acetate (pH5)500.0 ml iii)Adjust pH to 5.0 with 1.0 M NaOH c) 50 mM Ammonium acetate (pH5) i) Ammonium acetate.1.93 gm. ii) Distilled water500.0 ml iii)Adjust pH to 5.0 with 1.0 M HCl D) Treatment Procedure i) Rinse in PBS 2 times for 10 minutes each. ii) Rinse in distilled water 5 minutes. iii)Place slides in 10mM copper sulfate for 8 minutes. iv) Return slides to distilled water and check for autofluorescence with microscope. v) If needed return slides to 10mM copper sulfate for a couple of more minutes and check again. vi) Rinse slides for 5 minutes in distilled water. vii)Counterstain with DAPI 5 minutes. viii) Rinse in PBS 2 times for 10 minutes each. ix) Coverslip slides with appropriate mounting media. 3) 2.0 mM Glycine a) Used primarily for autofluorescence caused by free aldehyde groups. b) 2.0 mM Glycine i) Glycine...3.9 gm. ii) Distilled Water...26.0 ml c) Treatment Procedure i) Deparaffinize and rehydrate slides to H2O ii) Rinse in PBS 2 times for 10 minutes each. iii)Rinse in distilled water 5 minutes. iv) Place slides in 2.0 mM Glycine for 20-60 minutes. v) Rinse slides for 5 minutes in distilled water. vi) Rinse in PBS 2 times for 10 minutes each. vii)Continue with normal staining procedure 4) Sodium Borohydride a) The use of this reagent is particularly suited to reduce the reversible Schiff's bases that are formed by the aldehyde-NH2 reaction and lead to autofluorescence, especially when using glutaraldehyde. If you can use paraformaldehyde for fixation, the reduction step is often unnecessary and autofluorescence is low. This material has a high potential for explosion and is very caustic. b) The protocol was prepared by Jennifer Kramer and a similar procedure is described by Beisker, et al. (Beisker et al. 1987). i) Immediately before use (1) Make up a 1 mg/ml solution of sodium borohydride in a physiological buffer such as PBS. (a) The solution will be fizzy like carbonated water. Preparing this solution on ice and performing all subsequent incubations on ice has also been recommended. ii) Apply this solution immediately (while fizzing) to cells or tissue sections. (1) For glutaraldehyde fixed cell monolayers incubate in the sodium borohydride solution for 4 minutes. Replace with fresh sodium borohydride solution for another 4 minutes. (2) For paraformaldehyde fixed paraffin embedded 7 μm sections incubate 3 times, 10 minutes each in sodium borohydride solution. 5) Sudan Black a) This treatment is primarily for inhibition of autofluorescence in Lipfuscin. b) 0.3% Sudan Black (w/v) in 70% EtOH (v/v) stirred in the dark for 2 hours c) Apply to slide for 10 minutes after the secondary antibody application. d) Rinse quickly with PBS 8 times and mount e) For FITC and Alexa 594 this does not reduce the emission signal noticeably James Watson HT ASCP GNF Genomics Institute of the Novartis Research Foundation Tel 858-332-4647 Fax 858-812-1915 jwat...@gnf.org -Original Message- From:
[Histonet] North FL Histology Supervisor Job South FL Histology Supervisor Job
Hello, I currently have two available permanent positions open for a Histology Supervisor. One is in North Florida and one is in South Florida. Please let me know if you would like more details. Thank you, To view a complete list of Allied Search Partners current openings go to: http://www.alliedsearchpartners.com/careers.php -- Melissa Phelan President, Laboratory Staffing Allied Search Partners www.linkedin.com/in/melissaphelan/ http://www.linkedin.com/in/melissaphelan/ http://www.alliedsearchpartners.com http://www.alliedsearchpartners.com/ T: 888.388.7571 ext. 102 F: 888.388.7572 *If you wish to no longer receive emails from Allied Search Partners please respond to this email message with remove. This email including its attachments is intended only for the confidential use of the individual to whom it is addressed. If you are not the intended recipient, any use, dissemination, distribution or copying of this message or its attachments is prohibited. If you have received this message in error, please notify us immediately, and delete this message and its attachments permanently from your system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Manual IHC
Hi, I have been doing manual IHC for overflow of what doesn't fit in the Autostainer or for when I do not have enough to really justify running it (10 slides or so). Recently, I have been doing it exclusively since my Autostainer went belly up. Given the numbers you provided I assume you are doing about 75-80 slides per day. While I think this is entirely possible to do, you will find yourself dedicating pretty much the whole day to running these sliked and not having much time for anything else. Advantages: More direct control over each slide, More control over reagent volumes etc. Equipment costs. Disadvantages: It's really easy to get things mixed up and put the wrong reagent on the wrong slide unless you have a really good OCD eye to detail. Time variability (how long to rinse the first to last slide). Some tips if you do this: the Shandon Sequenza is great for applying reagents without fussing over keeping everything level. Rinse the slides *really* well. Be careful with the DAB since you don't need to worry much about it when using an instrument. My personal opinion on this is that given the number of slides you seem to be running an instrument is certainly justified. Best of luck, Amos Brooks ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Cytology Procedure
Hello all, I am looking for some assistance in Cytology procedures, specifically for non-gyn cytology. I am not as versed in Cytology as I am in Histology, so I am looking for some help with any procedures I might need. I know this is histonet, but I am sure a number of you have had to do cytology from time to time. Thanks in advance for any help. Haley Huggins, HT(ASCP)cm Pathology/Histology Manager Marian Medical Center 1400 East Church St Santa Maria, CA 93454 805-739-3170 (path lab) 805-739-3153 (office) 303-652-7453 (cell) 805-332-8697 (rightfax) ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] RE: Tissue marking dyes
Thank you all so much for your help and suggestions. I want to apologize for singling out a single company. It turns out, upon further examination, that we have inks from at least three different companies. TBS is actually the least problematic of the whole bunch. I won't mention who makes the others. Bill -Original Message- From: histonet-boun...@lists.utsouthwestern.edu [mailto:histonet-boun...@lists.utsouthwestern.edu] On Behalf Of O'Donnell, Bill Sent: Wednesday, October 16, 2013 10:51 AM To: Histonet (histonet@lists.utsouthwestern.edu) Subject: [Histonet] Tissue marking dyes I am wanting to 'cut' the dense tissue dye(s) by TBS (probably same as Cancer Diagnostics or any other). Orange is very thick, so is green. I'm being a bit lazy here, but I was wondering if anyone else is going this? Do you use water, alcohol or H2O2? Any direction would be helpful. Thanks - Bill William (Bill) O'Donnell, HT (ASCP) QIHC Senior Histologist Good Samaritan Hospital 10 East 31st Street Kearney, NE 68847 SERENITY is not freedom from the storm, but peace amid the storm. Cultivate it in PRAYER! This electronic mail and any attached documents are intended solely for the named addressee(s) and contain confidential information. If you are not an addressee, or responsible for delivering this email to an addressee, you have received this email in error and are notified that reading, copying, or disclosing this email is prohibited. If you received this email in error, immediately reply to the sender and delete the message completely from your computer system. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Desperately seeking information!!!
Hello all, A few weeks ago I sent out an information seeking email regarding IHC turnaround time. I did not get much in the way of responses. I figured there was not enough information provided to answer the general questions I was asking. I am having trouble obtaining an national average for IHC turnaround time. I am wondering if all you fellow histoneters out there would be willing to give me some info so I can see were we stand in comparison to facilities of similar size. The facility I work at turns out anywhere from 90-150 IHC stained slides daily. We have an average of 160 cases with around 700 HE stained slides daily. I have listed a few questions below, if any of you would be so kind as to take the time to answer them it would be greatly appreciated. What is the rough estimate of cases and initial HE stained slides that are turned out daily? Roughly, how many IHC stained slides do you turn out in a day? On average, what is your IHC turnaround time? What tissues are you working with (general surgical, dermatology's, research etc)? How many techs do you have that can perform IHC staining? Who is your instrumentation through? At the end of the day/run, is there a stain log printed? If so, who signs off on the positive/negative? If there are any other processes/procedures you feel are imperative to your IHC turnaround time please feel free to comment or offer suggestions. Thank you for taking the time to help us to improve our processes. If you have any questions or concerns please let me know. Again, thank you for your help, Emily K. McKenzie BS, HT(ASCP) Memorial Medical Center│701 North First Street│Springfield, IL 62781 Ph: 217-788-3991│email: mckenzie.em...@mhsil.com This message (including any attachments) contains confidential information intended for a specific individual and purpose, and is protected by law. If you are not the intended recipient, you should delete this message. Any disclosure, copying, or distribution of this message, or the taking of any action based on it, is strictly prohibited. ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
[Histonet] Diamond Knives
Good Evening, I would appreciate getting feedback on what you think of the different brands of Diamond Knives, pro's and con's quality, service, delivery, warranty, price, ... Thank you, Philip ___ Histonet mailing list Histonet@lists.utsouthwestern.edu http://lists.utsouthwestern.edu/mailman/listinfo/histonet
